Single-step multicolor fluorescence in situ hybridization using semiconductor quantum dot-DNA conjugates
Single-step multicolor fluorescence in situ hybridization using semiconductor quantum dot-DNA conjugates
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DOI：
10.1385/cbb:45:1:59
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发表时间：
2006-01-01
影响因子：
2.6
通讯作者：
Weiss, S
中科院分区：
文献类型：
--
作者：
Bentolila, LA;Weiss, S
我们报告了一种使用新型量子点荧光原位杂交（FISH）探针（QD-FISH）对多个亚核基因序列进行直接多色成像的快速方法。短DNA寡核苷酸附着在量子点上，用于原位杂交/检测目标位点的单一步骤。QD-FISH探针可以穿透完整的间期细胞核和中期染色体，并以最小的空间位阻很好地靶向密集的染色质结构域。我们进一步证明，量子点的宽吸收光谱允许不同颜色的探针特定于不同的亚核基因序列在一个单一的激发波长下同时被激发，并且在一次曝光中成像没有色差。因此，这些结果表明QD-FISH探针在多色FISH应用中是非常有效的。这项工作还记录了使用QD-FISH探针检测到单分子的新可能性。
We report a rapid method for the direct multicolor imaging of multiple subnuclear genetic sequences using novel quantum dot-based fluorescence in situ hybridization (FISH) probes (QD-FISH). Short DNA oligonucleotides were attached on QDs and used in a single hybridization/detection step of target sites in situ. QD-FISH probes penetrate both intact interphase nuclei and metaphase chromosomes and showed good targeting of dense chromatin domains with minimal steric hindrances. We further demonstrated that QD's broad absorption spectra allowed different colored probes specific for distinct subnuclear genetic sequences to be simultaneously excited with a single excitation wavelength and imaged free of chromatic aberrations in a single exposure. Thus, these results demonstrate that QD-FISH probes are very effective in multicolor FISH applications. This work also documents new possibilities of using QD-FISH probes detection down to the single molecule.